6.1. Chemistry
All the reagents were obtained commercially and used without further purification. Oxirane 1 and (4-oxo-2-thioxo-3-thiazolidinyl)acetic acid (23) were purchased from Sigma-Aldrich. The purity of the compounds was checked by thin-layer chromatography using silica-gel 60 F254-coated Al plates (Merck, Rahway, NJ, USA) and spots were observed under UV light. 1H NMR and 13C NMR spectra were recorded on a Bruker Avance spectrometer (600 and 150 MHz, respectively, Bruker, Billerica, MA, USA) and Varian VXR-400 spectrometer (Varian, Berkeley, CA, USA) at 400 MHz and 101 MHz, respectively, at 298 K in CDCl3 or DMSO-d6 at a concentration of samples of 5–15 mmol, with TMS as an internal reference for 1H and 13C NMR spectra. The signal assignments of compounds L-310 and 31a (L-173) were performed using 2D spectra (DQF-COSY, 13C–1H HSQC, and 13C–1H HMBC); the chemical shifts are expressed in ppm (δ scale) using DMSO-d6 or CDCl3 as an internal standard; the coupling constants expressed in Hz. The mass-spectral measurements were carried out by the ESI method on micrOTOF-QII (Brucker Daltonics GmbH, Bremen, Germany). Analytical HPLC was performed on a Shimadzu LC-20AD system using Kromasil-100-5-C18 (Akzo-Nobel, Amsterdam, The Netherlands) column, 4.6 × 250 mm, temperature 20 °C, UV detection, mobile phase A—0.2% HCOONH4), mobile phase B-MeCN, (pH 7.4), fl-1ml/min. LC/MS was performed on Agilent 1100 LC/MSD (Agilent Technologies, Santa Clara, CA, USA), MSD Ionization Source: APCI or ESI, detectors: ELSD, SEDEX 85, DAD (UV) 200–400 nm TWC (Total Wave Chromatogram).
3-(2-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)thiazolidine-2,4-dione (24) (Method A1). 2-(2,4-Dioxothiazolidin-3-yl)acetic acid (11) (175 mg, 1.0 mmol) was refluxed in 5 mL of SOCl2 for 2 h, then all volatiles evaporated in vacuo and the residue used without purification. 2-(2,4-Difluorophenyl)-4-(piperazin-1-yl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol hydrochloride (4a) (300 mg, 0.83 mmol) was suspended in 10 mL of CH2Cl2, then triethylamine (253 mg, 2.50 mmol, 3 equiv) was added. The reaction mixture was cooled in an ice bath and a solution of freshly prepared (see above) 2-(2,4-dioxothiazolidin-3-yl)acetyl chloride (194 mg, 1.0 mmol, 1.2 equiv) in CH2Cl2 was added dropwise. Upon completion of the acyl chloride addition, the pH of the mixture was adjusted to 8 with Et3N if needed. The reaction mixture was allowed to heat to room temperature and stirred overnight. Then it was washed with a saturated solution of citric acid until acidic pH, the organic layer was separated, dried over sodium sulfate and methylene chloride was distilled under vacuum. The resulting residue was redissolved in methylene chloride and the substance was purified by chromatography on silica gel (eluent—methanol:CH2Cl2 1:33). Yield 52%, light brown foam.
(Method A2) In methylene chloride (3 mL), 2-(2,4-dioxothiazolidin-3-yl)acetic acid (11) (0.42 mmol) and 2-(2,4-difluorophenyl)-1-(piperazin-1-yl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol (4a) hydrochloride (181 mg, 0.50 mmol, 1.2 equiv) were suspended, then triethylamine (169 mg, 1.67 mmol, 4 equiv) was added. The resulting solution was stirred for 30 min, and TBTU (161 mg, 0.50 mmol, 1.2 equiv) was added. pH was then adjusted to 8 with Et3N and the reaction mixture stirred at RT for 16 h. The organic layer was washed with water, dried over Na2SO4 and evaporated on a rotary evaporator. The residue was purified by flash chromatography on silica gel (eluent—ethyl acetate:hexane 3:1) to provide 24. Yield 32%, white foam.
1H NMR (400 MHz, DMSO-d6) δ 8.28 (s, 1H), 7.73 (s, 1H), 7.40 (td, J = 9.4, 9.0, 6.9, 1H), 7.13 (ddt, J = 11.5, 9.1, 2.4, 1H), 6.95 (td, J = 8.6, 2.5, 1H), 5.72 (s, 1H), 4.57 (d, J = 4.0, 2H), 4.33 (s, 1H), 4.26 (s, 1H), 3.98 (s, 1H), 3.46–3.42 (m, 4H + H2O), 2.86 (dd, J = 14.1, 7.2, 1H), 2.72–2.69 (dd, J = 13.9, 10.5, 1H), 2.38 (tp, J = 16.9, 5.3, 3H). 13C NMR (101 MHz, DMSO-d6) δ 172.23, 171.93, 163.37, 150.79, 145.25, 130.17, 111.09 (d, J = 20.6), 104.09 (t, J = 27.0), 75.04, 63.73, 55.94, 54.38, 54.09, 46.02, 44.46, 42.57, 42.11, 34.21, 21.44. LC/MS (ESI): m/z 481.1. Calculated for C20H22F2N6O4S [M + H]+ 481.1.
3-(1-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-1-oxopropan-2-yl)thiazolidin-2,4-dione (25) was obtained as described above from 12 and 4a. Yield 25%, crystalline yellow foam. 1H NMR (400 MHz, DMSO-d6) δ 8.26 (s, 1H), 7.72 (s, 1H), 7.38 (td, J = 8.9, 6.7, 1H), 7.12 (ddd, J = 11.9, 9.1, 2.6, 1H), 6.93 (td, J = 8.5, 2.6, 1H), 5.69 (s, 1H), 4.94 (qd, J = 7.1, 1.8, 1H), 4.55 (s, 2H), 4.21 (d, J = 2.7, 2H), 3.10 (s, 2H), 2.84 (d, J = 13.9, 1H), 2.67 (dd, J = 13.9, 2.1, 1H), 2.34 (s, 4H), 1.44–1.30 (m, 3H). 13C NMR (101 MHz, DMSO-d6) δ 171.95, 171.78, 166.34, 150.80, 145.24, 130.15, 111.09 (d, J = 20.2), 104.10 (t, J = 27.4), 75.10, 63.71, 55.94, 54.29, 49.97, 45.18, 42.41, 33.79, 14.79. LC/MS (ESI): m/z 495.2. Calculated for C21H24F2N6O4S [M + H]+ 495.2.
3-(4-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-4-oxobutyl)thiazolidine-2,4-dione (26) was obtained as described above from 13 and 4a. Yield 69%, yellow foam. 1H NMR (700 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.75 (s, 1H), 7.43–7.39 (m, 1H), 7.18–7.13 (m, 1H), 6.97 (td, J = 8.5, 2.5, 1H), 5.71 (s, 1H), 4.58 (s, 2H), 4.36 (t, J = 5.1, 1H), 4.15–4.12 (m, 2H), 3.49 (t, J = 6.9, 2H), 3.47–3.42 (m, 3H), 3.34–3.29 (m, 2H), 3.24 (t, J = 4.7, 2H), 2.87 (dd, J = 13.9, 0.9, 1H), 2.69 (d, J = 13.9, 1H), 2.51 (dt, J = 3.6, 1.8, 1H), 2.43–2.38 (m, 1H), 2.39–2.33 (m, 2H), 2.25 (t, J = 7.2, 2H), 1.70 (p, J = 7.1, 2H). 13C NMR (176 MHz, DMSO-d6) δ 172.91, 172.55, 169.96, 162.13 (dd, J = 245.7, 12.6), 159.41 (dd, J = 246.9, 12.1), 150.92, 145.37, 130.36–130.13 (m), 126.68–126.39 (m), 111.20 (d, J = 20.3), 104.21 (t, J = 26.9), 75.13 (d, J = 5.3), 63.90, 56.50, 56.06, 56.05, 54.54 (d, J = 38.3), 45.22, 41.45 (d, J = 45.7), 34.36, 29.72, 22.99, 19.01. LC/MS (ESI): m/z 509.2. Calculated for C22H26F2N6O4S [M + H]+ 509.2.
3-(5-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-4-oxopentyl)thiazolidine-2,4-dione (27) was obtained as described above from 14 and 4a. Yield 65%, white foam. 1H NMR (700 MHz, DMSO-d6) δ = 8.31 (s, 1H), 7.75 (s, 1H), 7.44–7.37 (m, 1H), 7.16 (t, J = 9.5, 1H), 7.00–6.94 (m, 1H), 4.58 (s, 2H), 4.19 (d, J = 7.1, 4H), 3.48 (dt, J = 12.6, 6.9, 5H), 2.85 (d, J = 14.0, 1H), 2.68 (d, J = 13.9, 1H), 2.30 (s, 2H), 1.90–1.72 (m, 3H). 13C NMR (176 MHz, DMSO-d6) δ 172.89, 172.87, 172.53, 172.51, 170.56, 162.15 (dd, J = 245.5, 12.7), 159.40 (dd, J = 247.0, 12.0), 150.93, 145.40, 130.26, 126.47, 111.23 (d, J = 20.1), 75.09, 63.84, 56.01, 54.80, 54.45, 41.40, 41.27, 40.30, 40.18, 40.07, 34.33, 34.32, 33.49, 31.98, 27.14, 26.95, 22.33, 22.06. LC/MS (ESI): m/z 523.2. Calculated for C23H28F2N6O4S [M + H]+ 523.2.
3-(1-(2,4-Dichlorophenyl)-2-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)thiazolidine-2,4-dione (28) was obtained from 15 and 4a. Yield 34%, yellow foam. 1H NMR (CDCl3) δ = 8.07 (s, 1H), 7.80 (s, 1H), 7.58–7.39 (m, 2H), 7.31 (m, 1H), 6.84 (m, 2H), 6.38–6.25 (m, 1H), 6.00–5.83 (m, 1H), 4.88 (br, 1H), 4.57 (m, 2H), 4.05–3.88 (m, 2H), 3.80–2.24 (m, 8H). LC/MS (ESI): m/z 625.1. Calculated for C26H24Cl2F2N6O4S [M + H]+ 625.1.
3-(2-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)-2-thioxothiazolidin-4-one (29) was obtained as described above from 23 and 4a. Yield 20%, brown foam. 1H NMR (700 MHz, DMSO-d6) δ = 8.30 (s, 1H), 7.75 (s, 1H), 7.41 (s, 1H), 7.16 (s, 1H), 6.97 (s, 1H), 5.76 (s, 1H), 4.78–4.28 (m, 5H), 3.31 (s, 2H), 2.89 (s, 1H), 2.69 (s, 2H), 2.56–2.31 (m, 4H). 13C NMR (176 MHz, DMSO-d6) δ 203.42, 174.39, 162.87, 162.17 (d, J = 246.6), 159.41 (dd, J = 246.9, 11.3), 150.92, 145.38, 130.26, 126.49 (d, J = 12.0), 111.22 (d, J = 20.2), 104.39, 104.23 (t, J = 26.9), 104.08, 75.19, 63.82, 56.02, 54.59, 54.28, 45.39, 44.75, 42.28, 38.71, 36.31. LC/MS (ESI): m/z 497.1. Calculated for C20H22F2N6O3S2 [M + H]+ 497.1.
3-(2-(4-(2-(2,4-Dichlorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)-2-thioxothiazolidin-4-one (30) was obtained as described above from 23 and 4b. Yield 28%, light brown foam. 1H NMR (700 MHz, DMSO-d6) δ = 8.32 (s, 2H), 7.86 (s, 1H), 7.75 (s, 2H), 7.68 (dd, J = 15.4, 7.4, 1H), 7.66–7.61 (m, 1H), 7.60–7.49 (m, 5H), 7.33 (s, 3H), 5.85 (s, 2H), 4.89 (d, J = 14.3, 3H), 4.29 (d, J = 4.9, 2H), 3.82–3.68 (m, 5H), 2.96–2.70 (m, 3H), 2.51 (s, 4H), 1.59 (s, 3H), 1.47 (d, J = 3.4, 6H). 13C NMR (176 MHz, DMSO-d6) δ 193.60, 181.20, 167.59, 166.95, 150.99, 145.53, 136.26, 133.03, 132.80, 132.62, 132.22, 131.99, 131.21, 130.24, 130.09, 127.30, 123.29, 54.88, 54.33, 54.12, 48.92, 46.93, 45.65, 44.82, 25.69, 24.41. LC/MS (ESI): m/z 529.1. Calculated for C20H22Cl2N6O3S2 [M + H]+ 529.1.
(Z)-5-(4-Chlorobenzylidene)-3-(2-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl) piperazin-1-yl)-2-oxoethyl)thiazolidine-2,4-dione (31a, L-173)5 was obtained as described above from 22a and 4a. Yield 67%, white crystals.
(Method B) To a solution of (Z)-5-(4-chlorobenzylidene)-3-(2-oxo-2-(piperazin-1-yl)ethyl)thiazolidin-2,4-dione (3.65 g, 10 mmol) in a mixture of PhMe:NMP—20:1 (50 mL), 1-((2-(2,4-difluorophenyl)oxiran-2-yl)methyl)-1H-1,2,4-triazole (2.37 g, 10 mmol) was added, followed by Et3N (5 mL, 38 mmol). The reaction mixture was stirred at 100 °C for 12 h, then cooled to room temperature and evaporated in vacuo. The residue was purified by flash chromatography using an EtOAc:MeOH:Et3N (25:1:0.1) mixture as an eluent. Yield 3.0 g (51%), crystalline powder. M.p. 150.3 °C. 1H NMR (700 MHz, DMSO-d6) δ 8.29 (s, 1H), 7.94 (s, 1H), 7.74 (s, 1H), 7.65 (d, J = 8.0, 2H), 7.57 (d, J = 8.0, 2H), 7.40 (td, J = 8.9, 6.8, 1H), 7.16 (ddd, J = 11.8, 9.1, 2.6, 1H), 6.96 (td, J = 8.4, 2.6, 1H), 5.73 (s, 1H), 4.58 (s, 2H), 4.54 (s, 2H), 3.4–3.25 (m, 4H), 2.88 (dd, J = 13.8, 1.5, 1H), 2.71 (d, J = 13.8, 1H), 2.49–2.40 (m, 3H), 2.40–2.31 (m, 4H). 13C NMR: (151 MHz, DMSO-d6) δ 166.78, 165.15, 162.89, 161.68 (dd, J = 245.9, 12.4), 158.94 (dd, J = 247.0, 12.0), 150.46, 144.90, 135.41, 132.17, 131.79, 131.71, 130.02–129.66 (m), 129.45, 126.03 (dd, J = 13.1, 3.6), 121.70, 110.98–110.49 (m), 103.76 (dd, J = 28.1, 25.7), 74.73 (d, J = 5.5), 63.37 (d, J = 3.6), 55.57 (d, J = 4.9), 54.04, 53.74, 44.18, 42.66, 41.83. HRMS (ESI): Calcd for C27H25ClF2N6O4S [M + H]+ 603.1387. Found: m/z 603.1485.
(Z)-3-(2-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)-5-(4-fluorobenzylidene)thiazolidine-2,4-dione (31b) was obtained from 22b and 4a. Yield 72%, white crystals. 1H NMR (400 MHz, DMSO-d6) δ 8.29 (s, 1H), 7.94 (s, 1H), 7.75 (s, 1H), 7.69 (dd, J = 8.7, 5.4, 2H), 7.45–7.30 (m, 3H), 7.14 (ddd, J = 11.8, 9.1, 2.6, 1H), 6.95 (td, J = 8.5, 2.6, 1H), 5.76 (s, 1H), 4.58 (s, 2H), 4.54 (s, 2H), 3.45–3.35 (m, 3H), 3.34–3.23 (m, 3H), 2.88 (d, J = 13.9, 1H), 2.70 (d, J = 13.8, 1H), 2.56–2.33 (m, 5H). 13C NMR (101 MHz, DMSO-d6) δ 167.37, 165.67, 163.49 (d, J = 251.3), 163.34, 162.11 (dd, J = 246.0, 12.7), 159.37 (dd, J = 246.7, 11.9), 150.92, 145.37, 133.17, 133.09, 132.85, 130.23 (t), 129.92 (d, J = 3.2), 126.48 (d, J = 13.0), 121.07 (d, J = 2.4), 117.00 (d, J = 22.0), 111.20 (d, J = 20.7), 104.21 (t, J = 26.8), 75.16 (d, J = 5.5), 62.48, 56.00 (d), 44.60, 43.07, 42.25. LC/MS (ESI): m/z 587.2. Calcd for C27H25F3N6O4S [M + H]+ 587.2.
(Z)-3-(2-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)-5-(4-methoxybenzylidene)thiazolidine-2,4-dione (31c) was obtained from 22c and 4a; Yield 61%, bright yellow foam. 1H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.88 (s, 1H), 7.75 (s, 1H), 7.64–7.55 (m, 3H), 7.41 (td, J = 9.0, 6.8, 1H), 7.23–7.05 (m, 4H), 6.96 (td, J = 8.5, 2.7, 1H), 5.77 (s, 1H), 4.58 (s, 2H), 4.51 (d, J = 9.5, 3H), 3.82 (d, J = 2.4, 5H), 3.70 (s, 1H), 3.47–3.22 (m, 6H), 2.89 (d, J = 13.9, 1H), 2.71 (d, J = 13.8, 1H), 2.57–2.30 (m, 7H). 13C NMR (101 MHz, DMSO-d6) δ 167.25, 167.12, 165.38, 162.98, 161.26, 150.46, 144.91, 134.15, 133.44, 132.47, 132.33, 129.98–129.57 (m), 126.35–125.65 (m), 125.29, 117.64, 114.99, 110.75 (d, J = 21.1), 103.76 (t, J = 27.0), 74.71 (d, J = 5.3), 63.34, 55.51, 54.03, 53.73, 52.65, 44.13, 42.52, 42.01, 41.77. LC/MS (ESI): m/z 599.2. Calcd for C28H28F2N6O5S [M + H]+ 599.2.
(Z)-5-(4-Ethylbenzylidene)-3-(2-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)thiazolidine-2,4-dione (31d) was obtained from 22d and 4a. Yield 72%, white crystals. 1H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.90 (s, 1H), 7.75 (s, 1H), 7.54 (d, J = 8.1, 2H), 7.40 (s, 1H), 7.38–7.33 (m, 2H), 7.15 (ddd, J = 11.8, 9.1, 2.6, 1H), 6.96 (td, J = 8.5, 2.6, 1H), 5.76 (s, 1H), 4.58 (s, 2H), 4.53 (s, 2H), 3.36 (d, J = 4.9, 3H), 3.34–3.22 (m, 1H), 2.95–2.83 (m, 1H), 2.71 (d, J = 13.8, 1H), 2.64 (q, J = 7.6, 2H), 2.42 (qd, J = 12.8, 12.3, 6.3, 2H), 1.17 (t, J = 7.6, 3H). 13C NMR (101 MHz, DMSO-d6) δ 167.50, 165.74, 163.35, 160.86 (d, J = 12.3), 160.57 (d, J = 12.2), 158.12 (d, J = 12.3), 150.88, 147.65, 145.32, 133.94, 130.76 (d, J = 4.2), 130.17 (d, J = 8.6), 129.26, 126.45 (d, J = 11.7), 120.17, 111.16 (d, J = 20.4), 104.19 (t, J = 26.8), 75.14 (d, J = 5.4), 63.77, 55.96, 54.46, 54.16, 44.57, 42.99, 42.22, 28.55, 15.59. HRMS (ESI): Calcd for C29H30F2N6O4S [M + H]+ 597.2090. Found: m/z 597.2103.
(Z)-5-(4-Isopropylbenzylidene)-3-(2-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)thiazolidine-2,4-dione (31e) was obtained from 22e and 4a. Yield 65%, white crystals. 1H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.90 (s, 1H), 7.75 (s, 1H), 7.62–7.52 (m, 2H), 7.47–7.38 (m, 3H), 7.16 (ddd, J = 11.8, 9.1, 2.6, 1H), 6.96 (td, J = 8.5, 2.6, 1H), 4.58 (s, 2H), 4.54 (s, 2H), 3.37 (t, J = 5.0, 2H), 3.35–3.21 (m, 2H), 3.00–2.84 (m, 2H), 2.71 (d, J = 13.7, 1H), 2.41 (tt, J = 12.9, 6.7, 4H), 1.20 (d, J = 6.9, 6H). 13C NMR (101 MHz, DMSO-d6) δ 167.51, 165.74, 163.35, 160.85 (d, J = 12.5), 160.63, 158.05, 152.13, 150.88, 145.32, 133.90, 130.86 (d, J = 6.7), 130.16 (d, J = 9.4), 127.84, 126.44 (d, J = 13.7), 120.21, 111.17 (d, J = 20.1), 104.60–103.83 (m), 85.45, 75.12 (d, J = 5.5), 63.76, 55.95, 54.45, 54.15, 44.54, 42.99, 42.19, 33.86, 23.90. HRMS (ESI): Calcd for C30H32F2N6O4S [M + H]+ 611.2247. Found: m/z 612.2445.
(Z)-5-(4-tert-Butylbenzylidene)-3-(2-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)thiazolidine-2,4-dione (31f) was obtained from 22f and 4a. Yield 63%, white crystals. 1H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.91 (s, 1H), 7.75 (s, 1H), 7.57 (d, J = 3.0, 4H), 7.40 (td, J = 9.0, 6.8, 1H), 7.16 (ddd, J = 11.8, 9.1, 2.6, 1H), 6.96 (td, J = 8.5, 2.6, 1H), 5.78 (s, 1H), 4.58 (s, 2H), 4.54 (s, 2H), 3.37 (d, J = 5.2, 2H), 3.34–3.21 (m, 3H), 2.89 (d, J = 13.9, 1H), 2.71 (d, J = 13.8, 1H), 2.42 (tt, J = 12.9, 6.7, 4H), 1.28 (s, 9H). 13C NMR (101 MHz, DMSO-d6) δ 167.51, 165.74, 163.35, 160.85 (d, J = 12.5), 160.51, 158.12 (d, J = 12.0), 154.30, 150.88, 145.33, 133.78, 130.57, 130.22, 126.71, 126.59–126.28 (m), 120.32, 111.17 (d, J = 20.6), 104.19 (t, J = 26.9), 75.14 (d, J = 5.4), 63.78, 55.95, 54.46, 54.15, 44.57, 43.00, 42.21, 35.20, 31.19. HRMS (ESI):Calcd for C31H34F2N6O4S [M + H]+ 625.2403. Found: m/z 625.2295.
(Z)-5-(2,4-Dichlorobenzylidene)-3-(2-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)thiazolidine-2,4-dione (31g) was obtained from 22g and 4a. Yield 58%, white crystals. 1H NMR (400 MHz, DMSO-d6) δ 8.28 (s, J = 4.1, 1H), 7.94 (d, J = 8.3, 2H), 7.83 (s, 1H), 7.74 (s, 1H), 7.60 (s, 2H), 7.40 (dd, J = 15.9, 8.5, 1H), 7.14 (t, J = 9.7, 1H), 6.95 (t, J = 7.4, 1H), 5.73 (s, 1H), 4.57 (s, J = 10.3, 2H), 4.55 (s, 2H), 3.37 (s, 2H), 2.87 (s, 5H), 2.71 (s, 5H), 2.46–2.32 (m, 3H). 13C NMR (101 MHz, DMSO-d6) δ 167.00, 165.22, 163.23, 162.72, 150.90, 145.34, 136.28, 135.86, 130.60, 130.42, 130.34, 130.22, 128.84, 127.91, 125.76, 111.28, 111.07, 104.47, 104.19, 103.95, 75.19, 75.13, 63.77, 59.91, 56.02, 55.96, 54.48, 54.17, 49.97, 44.62, 43.20, 42.43, 42.28, 36.21, 31.20. LC/MS (ESI): m/z 637.1. Calculated for C27H24Cl2F2N6O4S [M + H]+ 637.1.
(Z)-5-(4-Chlorobenzylidene)-3-(1-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazine-1-yl)-1-oxopropan-2-yl)thiazolidine-2,4-dione (32a) was obtained from 23a and 4a. Yield 72%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.26 (s, 1H), 7.92–7.90 (m, 1H), 7.72 (s, 1H), 7.64 (d, J = 8.4, 2H), 7.60 (d, J = 8.3, 2H), 7.37 (q, J = 8.3, 1H), 7.16–7.09 (m, 1H), 6.93 (td, J = 8.4, 2.6, 1H), 5.69 (d, J = 2.5, 1H), 5.19 (qd, J = 7.2, 2.9, 1H), 4.55 (s, 2H), 3.35–3.30 (m, 3H), 3.20 (s, 2H), 2.83 (dd, J = 14.0, 4.9, 1H), 2.67 (dd, J = 14.0, 4.4, 1H), 2.45–2.40 (m, 3H), 2.33 (s, 2H), 2.29 (s, 1H), 1.49 (dd, J = 7.2, 4.0, 3H). 13C NMR (176 MHz, DMSO-d6) δ 166.82, 166.42, 165.46, 162.02 (dd, J = 245.7, 12.5), 159.29 (dd, J = 247.0, 12.0), 150.84, 145.27, 135.75, 132.51, 132.23, 132.18, 130.16, 129.87, 126.36 (d, J = 13.1), 121.62, 111.11 (d, J = 20.5), 104.13 (t, J = 26.9), 75.15, 63.71, 55.92, 54.45, 54.40, 54.27, 50.85 (d, J = 7.7), 45.34, 42.52, 14.90. LC/MS (ESI): m/z 617.2. Calculated for C28H27ClF2N6O4S [M + H]+ 617.2
The compound was also synthesized by Method B (see 31a) with a 46% yield.
(Z)-5-(4-Chlorobenzylidene)-3-(4-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-4-oxobutyl)thiazolidine-2,4-dione (33a) was obtained from 24a and 4a. Yield 41%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ = 8.30 (s, 1H), 7.89 (s, 1H), 7.75 (s, 1H), 7.63 (d, J = 8.6, 2H), 7.60 (d, J = 8.6, 1H), 7.40 (d, J = 7.0, 1H), 7.16–7.13 (m, 1H), 6.96 (td, J = 8.5, 2.4, 1H), 5.84–5.60 (m, 1H), 4.58 (s, 2H), 3.66 (br. s, 2H), 2.85 (d, J = 14.0, 1H), 2.68 (d, J = 13.9, 1H), 2.30 (s, 2H), 1.90–1.72 (m, 3H). 13C NMR (176 MHz, DMSO-d6) δ 169.93, 167.63, 166.12, 162.12 (dd, J = 245.7, 12.4), 159.39 (dd, J = 246.9, 12.2), 150.91, 145.37, 135.61, 132.39, 132.15, 131.62, 130.25, 129.91, 126.53 (d, J = 13.3), 122.83, 111.19 (d, J = 20.5), 104.20, 75.13 (d, J = 5.3), 63.90, 56.02, 54.62, 54.39, 45.22, 41.92, 41.60, 29.74, 22.96. LC/MS (ESI): m/z 631.2. Calculated for C29H29ClF2N6O4S [M + H]+ 631.2.
The compound was also synthesized by Method B (see 31a) with a 32% yield.
(Z)-3-(4-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-4-oxobutyl)-5-(4-fluorobenzylidene)thiazolidine-2,4-dione (33b) was obtained from 24b and 4a. Yield 37%, yellow crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.90 (s, 1H), 7.75 (s, 1H), 7.72–7.65 (m, 3H), 7.44–7.33 (m, 4H), 7.15 (t, J = 9.5, 1H), 6.96 (t, J = 7.2, 1H), 5.91–5.55 (m, 1H), 4.58 (s, 2H), 3.66 (t, J = 6.8, 2H), 3.26 (br. s, 6H), 2.47–2.21 (m, J = 7.0, 8H), 2.30 (t, J = 7.0, 2H), 1.82–1.79 (m, 2H). 13C NMR (176 MHz, DMSO-d6) δ 169.96, 167.77, 166.19, 163.40 (d, J = 251.0), 162.16 (dd, J = 245.7, 12.2), 159.39 (dd, J = 246.9, 12.1), 150.94, 145.41, 133.04, 132.99, 131.91, 130.25, 130.16, 126.43, 121.75, 117.02 (d, J = 22.0), 111.24 (d, J = 20.4), 104.24 (t, J = 26.8), 75.04, 56.01, 54.35, 41.87, 29.73, 22.97. LC/MS (ESI): m/z 615.2. Calculated for C29H29F3N6O4S [M + H]+ 615.2.
(Z)-5-(2,4-Dichlorobenzylidene)-3-(4-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-4-oxobutyl)thiazolidin-2,4-dione (33g) was obtained from 13g and 4a. Yield 45%, yellow crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.93 (s, 1H), 7.84 (d, J = 2.0, 1H), 7.75 (s, 1H), 7.61 (d, J = 1.9, 1H), 7.57 (d, J = 8.5, 1H), 7.41 (d, J = 7.0, 1H), 7.18–7.11 (m, J = 9.3, 1H), 6.97–6.95 (m, 1H), 5.72 (s, 1H), 4.58 (s, 2H), 3.67 (t, J = 6.8, 2H), 3.25 (s, 3H), 2.85 (d, J = 13.9, 1H), 2.68 (d, J = 13.9, 1H), 2.45–2.24 (m, 7H), 1.86–1.76 (m, 2H). 13C NMR (176 MHz, DMSO-d6) δ 169.91, 167.41, 165.79, 162.12 (dd, J = 245.8, 12.4), 159.39 (dd, J = 246.9, 12.1), 150.92, 145.37, 136.07, 135.85, 130.50, 130.48, 130.45, 130.26 (t), 128.86, 126.51 (d, J = 2.0), 111.19 (d, J = 20.1), 104.20, 75.13 (d, J = 5.1), 63.90, 56.04, 56.02, 54.62, 54.41, 45.22, 42.09, 41.61, 29.77, 25.80, 22.88. LC/MS (ESI): m/z 665.1. Calculated for C29H28Cl2F2N6O4S [M + H]+ 665.1.
(Z)-3-(2-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)-5-(4-chlorobenzylidene)-2-thio-1,3-thiazolidin-4-one (29a) (Method K): To a solution of 29 (0.33 mmol) in 3 mL of ethanol, 4-chlorobenzaldehyde (0.36 mmol, 1.1 equiv) was added, followed by piperidine (6 mg, 0.07 mmol, 0.2 equiv.). The mixture refluxed for 16 h, then was cooled and evaporated on a rotary evaporator. The product was isolated by flash chromatography on silica gel (elution system — 15% methanol in CH2Cl2). Yield 60%, yellow crystals. 1H NMR (700 MHz, DMSO-d6) δ = 8.36 (s, 1H), 7.89 (s, 1H), 7.81 (s, 1H), 7.72 (d, J = 8.2, 2H), 7.67 (d, J = 8.3, 2H), 7.47 (q, J = 8.4, 1H), 7.22 (ddd, J = 11.7, 8.9, 2.6, 1H), 7.03 (td, J = 8.5, 2.5, 1H), 5.82 (s, 1H), 4.95 (s, 2H), 4.65 (s, 1H), 3.53–3.47 (m, 3H), 3.38 (dt, J = 10.5, 5.2, 2H), 2.96 (d, J = 13.8, 1H), 2.54–2.40 (m, 2H). 13C NMR (176 MHz, DMSO-d6) δ 193.56, 166.97, 162.76, 162.16 (dd, J = 246.0, 12.7), 159.42 (dd, J = 247.0, 12.1), 150.95, 145.39, 136.27, 132.60, 132.20, 130.28 (t, J = 8.2), 130.07, 126.4 (d, J = 12.9), 123.26, 111.26 (d, J = 20.4), 104.25 (t, J = 27.0), 75.19 (d, J = 5.3), 63.82, 56.03 (d, J = 5.0), 54.61, 54.29, 45.67, 44.82, 42.35. HRMS (EI): Calcd for C27H25ClF2N6O3S2 [M + H]+ 619.1159. Found: 619.1137.
(Z)-5-(4-Chlorobenzylidene)-3-(2-(4-(2-(2,4-dichlorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)-2-thioxothiazolidin-4-one (30a) was obtained as described above from 30 and 4-chlorobenzaldehyde. Yield 58%, bright yellow crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.32 (s, 1H), 7.84 (s, 1H), 7.76 (s, 1H), 7.68 (d, J = 8.4, 2H), 7.62 (d, J = 8.3, 2H), 7.59–7.51 (m, 2H), 7.33 (d, J = 7.8, 1H), 5.86 (s, 1H), 4.91 (s, 1H), 4.89 (s, 2H), 4.67 (d, J = 14.0, 1H), 3.33–3.20 (m, 3H), 2.82 (d, J = 10.3, 1H), 2.47–2.24 (m, 3H). 13C NMR (176 MHz, DMSO-d6) δ 193.56, 166.95, 162.76, 150.98, 145.51, 136.26, 133.03, 132.79, 132.61, 132.20, 132.02, 131.22, 130.22, 130.08, 127.28, 123.26, 76.18, 62.31, 54.89, 54.32, 54.04, 45.65, 44.73, 42.27. LC/MS (ESI): m/z 651.1. Calculated for C27H25Cl3N6O3S2 [M + H]+ 651.1.
(Z)-5-(4-Methoxybenzylidene)-3-(2-(4-(2-(2,4-dichlorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)-2-thio-1,3-thiazolidin-4-one (30c) was obtained as described above from 30 and 4-methoxybenzaldehyde. Yield 39%, bright yellow crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.32 (s, 1H), 7.80 (s, 1H), 7.76 (s, 1H), 7.62 (d, J = 8.7, 2H), 7.57 (d, J = 8.6, 1H), 7.54 (s, 1H), 7.33 (d, J = 8.1, 1H), 7.12 (d, J = 8.7, 2H), 5.87 (s, 1H), 4.90 (d, J = 14.3, 1H), 4.88 (s, 2H), 4.67 (d, J = 14.3, 1H), 3.84 (s, 3H), 3.34–3.18 (m, 1H), 2.82 (d, J = 11.1, 1H), 2.48–2.28 (m, 1H). 13C NMR (176 MHz, DMSO-d6) δ 193.68, 167.09, 162.87, 162.14, 150.97, 145.50, 134.20, 133.51, 133.03, 132.02, 131.22, 130.22, 127.28, 125.88, 119.17, 115.66, 76.17, 62.31, 56.07, 54.89, 54.32, 54.04, 45.57, 44.73, 42.25. LC/MS (ESI): m/z 647.1. Calculated for C28H28Cl2N6O4S2 [M + H]+ 647.1.
(Z)-5-(3,5-Dichloro-2-hydroxybenzylidene)-3-(5-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazole-1-yl)propyl)piperazin-1-yl)-5-oxobutyl)thiazolidine-2,4-dione (33h) was obtained as described above from 26 and 3,5-dichloro-2-hydroxybenzaldehyde. Yield 51%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.98 (s, 1H), 7.75 (d, J = 3.4, 1H), 7.65 (d, J = 2.2, 1H), 7.40 (q, J = 8.8, 1H), 7.27 (d, J = 2.2, 1H), 7.19–7.10 (m, 1H), 6.96 (dd, J = 8.1, 6.5, 1H), 4.58 (s, 2H), 3.66 (t, J = 6.8, 2H), 2.86 (s, 1H), 2.69 (d, J = 13.8, 1H), 2.46–2.32 (m, 4H), 1.85–1.73 (m, 2H). 13C NMR (176 MHz, DMSO-d6) δ 192.49, 172.92, 172.55, 169.98, 169.93, 167.60, 166.00, 162.13 (dd, J = 245.7, 12.3), 159.39 (dd, J = 247.1, 12.1), 152.23, 150.92, 145.37, 135.49, 131.35, 130.24 (t), 129.07, 126.95, 126.51 (d, J = 12.6), 124.90, 124.21, 123.92, 123.81, 111.20 (d, J = 20.4), 104.21 (t, J = 26.7), 75.08 (d, J = 4.9), 63.88, 56.04, 54.62, 54.40, 45.19, 41.90, 41.57, 41.32, 34.36, 29.75, 23.00. LC/MS (ESI): m/z 681.1. Calculated for C29H28Cl2F2N6O5S [M + H]+ 681.1.
(Z)-5-(2-Chloro-6-(4-methylpiperazin-1-yl)benzylidene)-3-(4-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-4-oxobutyl)thiazolidine-2,4-dione (33i) was obtained as described above from 26 and 2-chloro-6-(4-methylpiperazino)benzaldehyde. Yield 46%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.00 (s, 1H), 7.80 (s, 1H), 7.50–7.42 (m, J = 8.1, 2H), 7.27 (d, J = 7.9, 1H), 7.21 (d, J = 8.1, 1H), 7.20–7.17 (m, 1H), 7.01 (td, J = 8.5, 2.5, 1H), 5.82 (s, 1H), 4.63 (s, 2H), 3.70 (t, J = 6.8, 2H), 3.05 (br.s, 5H), 2.92 (d, J = 14.1, 1H), 2.73 (d, J = 13.8, 1H), 2.66 (s, 4H), 2.50–2.30 (m, 9H), 1.90–1.82 (m, 2H). 13C NMR (176 MHz, DMSO-d6) δ 169.88, 167.87, 166.04, 163.61, 162.11 (dd, J = 245.7, 12.2), 159.40 (dd, J = 246.9, 12.0), 152.53, 150.90, 145.39, 134.34, 132.08, 130.26, 129.93, 126.56 (d, J = 12.8), 126.04, 125.18, 124.00, 118.47, 111.19 (d, J = 20.2), 104.18 (t, J = 26.9), 75.15 (d, J = 5.4), 63.90, 56.02, 54.60, 54.39, 54.17, 51.24, 45.25, 44.05, 41.75, 41.62, 29.83, 23.03, 22.59. LC/MS (ESI): m/z 729.3. Calculated for C34H39ClF2N8O4S [M + H]+ 729.3.
(Z)-5-(4-Chlorobenzylidene)-3-(5-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-5-oxopentyl)thiazolidin-2,4-dione (34a) was obtained as described above from 27 and 4-chlorobenzaldehyde. Yield 44%, yellow crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.90 (s, 1H), 7.75 (s, 1H), 7.63 (d, J = 8.4, 2H), 7.59 (d, J = 8.5, 2H), 7.45–7.35 (m, 1H), 7.13 (t, J = 9.3, 1H), 6.95 (t, J = 7.2, 1H), 4.57 (s, J = 7.5, 2H), 3.64 (t, J = 6.9, 2H), 2.85 (d, J = 14.0, 1H), 2.68 (d, J = 13.8, 1H), 2.44–2.30 (m, 4H), 2.30–2.19 (m, 2H), 1.70–1.52 (m, 2H), 1.52–1.36 (m, 2H). 13C NMR (176 MHz, DMSO-d6) δ 174.64, 170.52, 167.54, 166.03, 162.13 (dd, J = 245.8, 12.3), 159.38 (dd, J = 247.0, 12.1), 150.91, 145.37, 135.69, 132.32, 132.18, 131.98, 130.24 (t), 129.89, 126.50 (d, J = 12.9), 122.54, 111.19 (d, J = 20.3), 104.20 (t, J = 26.9), 75.11 (d, J = 5.1), 63.87, 56.02, 54.84, 54.48, 45.32, 41.93, 41.49, 32.00, 27.22, 22.4. LC/MS (ESI): m/z 645.2. Calculated for C30H31ClF2N6O4S [M + H]+ 645.2.
(Z)-3-(5-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-5-oxopentyl)-5-(4-fluorobenzylidene)thiazolidine-2,4-dione (34b) was obtained from 27 and 4-fluorobenzaldehyde. Yield 48%, yellow crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.94 (s, 1H), 7.75 (s, 1H), 7.73–7.67 (m, 2H), 7.48–7.33 (m, 3H), 7.18–7.13 (m, 1H), 7.00–6.91 (m, 1H), 4.57 (s, 2H), 3.64 (t, J = 6.9, 2H), 3.29 (s, 4H), 2.87 (d, J = 12.2, 1H), 2.76–2.61 (m, 1H), 2.37 (s, 4H), 2.30–2.16 (m, 3H), 1.69–1.34 (m, 6H). 13C NMR (176 MHz, DMSO-d6) δ 174.63, 170.52, 167.69, 166.10, 163.44 (d, J = 250.8), 162.14 (dd, J = 245.8, 12.3), 159.39 (dd, J = 247.0, 12.1), 150.93, 145.39, 133.07 (d, J = 8.8), 132.26, 130.25 (t), 130.12, 126.46 (d, J = 11.9), 121.50, 117.02 (d, J = 22.0), 116.08 (d, J = 22.3), 111.22 (d, J = 20.2), 104.23 (t, J = 26.9), 75.06, 63.83, 56.01, 54.81, 54.46, 45.25, 41.89, 41.40, 33.46, 31.99, 27.23, 27.15, 27.06, 22.41, 22.33, 22.11. LC/MS (ESI): m/z 629.2. Calculated for C30H31F3N6O4S [M + H]+ 629.2.
(Z)-5-(2,4-Dichlorobenzylidene)-3-(5-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-5-oxopentyl)thiazolidin-2,4-dione (34g) was obtained from 27 and 2,4-dichlorobenzaldehyde. Yield 44%, bright yellow crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.94 (s, 1H), 7.82 (d, J = 1.6, 1H), 7.74 (s, 1H), 7.67–7.54 (m, 2H), 7.46–7.36 (m, 1H), 7.19–7.09 (m, 1H), 6.95 (t, J = 6.8, 1H), 4.58 (s, 2H), 3.64 (t, J = 6.9, 2H), 3.44–3.18 (m, 6H), 2.85 (d, J = 14.2, 1H), 2.68 (d, J = 13.7, 1H), 2.46–2.30 (m, 4H), 2.30–2.21 (m, 2H), 1.60–1.55 (m, 2H), 1.52–1.31 (m, 4H). 13C NMR (176 MHz, DMSO-d6) δ 174.62, 170.49, 167.34, 165.70, 162.12 (dd, J = 245.7, 12.4), 159.39 (dd, J = 246.9, 12.1), 150.91, 145.37, 136.14, 135.89, 130.53, 130.43, 130.25, 130.20, 128.83, 127.07, 126.52 (d, J = 12.9), 126.14, 111.19 (d, J = 20.3), 104.19 (t, J = 26.9), 75.12 (d, J = 4.7), 63.89, 56.02, 54.85, 54.49, 45.34, 42.09, 41.50, 32.00, 27.19, 22.41, 22.12. LC/MS (ESI): m/z 679.1. Calculated for C30H30Cl2F2N6O4S [M + H]+ 679.1.
(Z)-5-(3,5-Dichloro-2-hydroxybenzylidene)-3-(5-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-5-oxopentyl)thiazolidine-2,4-dione (34h) was obtained as described above from 27 and 3,5-dichloro-2-hydroxybenzaldehyde. Yield 41%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.30 (d, J = 2.8, 1H), 7.99 (s, 1H), 7.75 (d, J = 4.3, 1H), 7.65 (d, J = 2.4, 1H), 7.45–7.35 (m, 1H), 7.31–7.25 (m, 1H), 7.18–7.10 (m, 1H), 6.96 (dd, J = 5.9, 2.4, 1H), 4.58 (d, J = 5.0, 2H), 4.18 (d, J = 7.3, 1H), 2.87 (t, J = 13.8, 1H), 2.73–2.64 (m, 1H), 2.47–2.30 (m, 4H), 2.29–2.16 (m, 2H), 1.65–1.31 (m, 6H). 13C NMR (176 MHz, DMSO-d6) δ 174.65, 172.85, 172.50, 170.54, 167.54, 165.94, 162.13 (dd, J = 245.7, 12.4), 159.39 (dd, J = 247.5, 12.0), 152.21, 150.92, 145.37, 131.40, 130.23, 127.30, 127.04, 126.53, 126.49 (d, J = 13.0), 124.85, 124.02, 123.94, 123.81, 111.21 (d, J = 20.2), 104.29 (d, J = 26.3), 75.09 (d, J = 4.8), 63.86, 56.03, 54.83, 54.48, 45.30, 41.93, 41.47, 41.40, 41.27, 34.31, 33.49, 33.46, 31.99, 27.22, 27.15, 27.05, 26.95, 22.41, 22.34, 22.11, 22.07. LC/MS (ESI): m/z 695.1. Calculated for C30H30Cl2F2N6O5S [M + H]+ 695.1.
(Z)-5-(2-Chloro-6-(4-methylpiperazin-1-yl)benzylidene)-3-(5-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-5-oxopentyl)thiazolidin-2,4-dione (34i) was obtained as described above from 27 and 2-chloro-6-(4-methylpiperazin-1-yl)benzaldehyde. Yield 38%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ = 8.30 (s, 1H), 7.98 (s, 1H), 7.75 (s, J = 3.6, 1H), 7.40 (t, J = 8.0, 2H), 7.20 (d, J = 7.9, 1H), 7.14 (d, J = 8.3, 2H), 7.00–6.91 (m, 1H), 5.73 (s, 1H), 4.57 (s, 2H), 3.64 (t, J = 6.8, 2H), 3.59–3.10 (m, 10H), 2.94 (s, 3H), 2.85 (d, J = 13.7, 2H), 2.67 (d, J = 13.9, 1H), 2.44 (s, 3H), 2.42–2.29 (m, 5H), 2.27 (t, J = 7.2, 2H), 2.20 (s, 4H), 1.63–1.53 (m, 2H), 1.46–1.38 (m, 2H). 13C NMR (176 MHz, DMSO-d6) δ 170.48, 167.77, 165.98, 163.51, 162.12 (dd, J = 245.6, 12.3), 159.39 (dd, J = 246.8, 12.3), 152.79, 150.91, 145.37, 134.35, 132.11, 130.41, 130.24, 126.54 (d, J = 12.6), 125.62, 125.05, 123.79, 118.42, 111.19 (d, J = 20.3), 104.20 (t, J = 26.8), 75.14 (d, J = 5.2), 63.90, 56.04, 55.31, 54.86, 54.55, 54.49, 51.82, 45.96, 45.34, 41.69, 41.51, 31.98, 27.31, 22.38. LC/MS (ESI): m/z 743.3. Calculated for C35H41ClF2N8O4S [M + H]+ 743.3.
(Z)-3-(1-(2,4-Dichlorophenyl)-2-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-2-oxoethyl)-5-(4-fluorobenzylidene)thiazolidine-2,4-dione (35b) was obtained from 30 and 4-fluorobenzaldehyde. Yield 33%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.27 (d, J = 1.1, 1H), 7.95 (s, 1H), 7.74 (d, J = 1.2, 1H), 7.72–7.68 (m, 3H), 7.52–7.47 (m, 1H), 7.43–7.35 (m, 4H), 7.13 (t, J = 9.8, 1H), 6.94 (td, J = 8.5, 2.5, 1H), 6.32 (d, J = 2.3, 1H), 5.70 (s, 1H), 4.55 (d, J = 6.7, 2H), 3.47 (d, J = 23.1, 2H), 3.09 (d, J = 3.2, 1H), 2.91–2.76 (m, 2H), 2.68 (t, J = 13.8, 1H), 2.59–2.37 (m, 3H), 2.36–2.15 (m, 2H). 13C NMR (176 MHz, DMSO-d6) δ 166.62, 165.09, 163.62 (d, J = 251.5), 163.10 (d, J = 4.0), 163.11, 163.09, 162.11 (dd, J = 245.8, 12.5), 159.35 (dd, J = 248.7, 10.2), 150.94, 145.36, 135.06, 134.37, 134.18, 134.14, 133.78, 133.34, 133.29, 130.24, 129.88, 129.65, 129.10, 129.07, 127.63, 126.36 (d, J = 11.8), 119.80, 117.14, 117.02, 111.21 (d, J = 20.4), 104.23 (t, J = 26.8), 75.18 (dd, J = 12.6, 5.2), 63.67, 63.61, 56.54, 56.03, 55.95, 54.05, 54.00, 53.91, 45.80, 42.76. HRMS (EI): Calcd for C33H27Cl2F3N6O4S [M + H]+ 731.1216. Found: m/z 731.1033.
3-Chloro-1-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)-piperazin-1-yl)propan-1-one (49). 2-(2,4-Difluorophenyl)-1-(piperazin-1-yl)-3-(1H-1,2,4-triazol-1-yl)propan-2-ol hydrochloride (4b) (700 mg, 1.95 mmol) was suspended in methylene chloride (20 mL), then triethylamine (591 mg, 5.84 mmol, 3 equiv) was added. RM was cooled in an ice bath, and 3-chloropropionyl chloride (296 mg, 2.33 mmol, 1.2 equiv) was added dropwise. Upon full addition of the acyl chloride, the pH of the mixture was adjusted to 8 with Et3N if needed. The reaction mixture was allowed to heat to room temperature and stirred overnight. It was then washed with a saturated citric acid solution to achieve an acidic pH, the organic layer was separated, dried over Na2SO4, and the solvent evaporated under vacuum. The resulting residue was redissolved in CH2Cl2 and purified by flash chromatography on silica gel (eluent—methanol:CH2Cl2 1:25). Yield 80% (647 mg), dark yellow oil. The substance was used in the next step without purification. LC/MS(+ESI): Found m/z 414.2. Calcd for C18H22ClF2N5O2 [M + H]+ 414.2.
(Z)-5-(4-Chlorobenzylidene)-3-(3-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-3-oxopropyl)thiazolidin-2,4-dione (50a) (Method C). Potassium salt of 5-(4-chlorobenzylidene)thiazolidine-2,4-dione (222 mg, 0.8 mmol) is suspended in 4 mL of DMF, then 4-(3-chloroethylcarbonylpiperazin-1-yl)-3-(1H-1,2,4-triazolyl-1-yl)propan-2-ol (49) (322 mg, 0.8 mmol, 1 equiv.) and potassium iodide (134 mg, 0.8 mmol, 1 equiv) were added. The resulting suspension was stirred for 12 h at 80 °C. Then, the reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (3 × 3 mL). The organic layer was separated, dried over Na2SO4, and evaporated on a rotary evaporator. The residue was redissolved in CH2Cl2, and subjected to chromatography on silica gel (eluent— 5% methanol in methylene chloride). Yield 18%, pale yellow foam. 1H NMR (700 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.93 (s, 1H), 7.75 (s, 1H), 7.65 (d, J = 8.7, 2H), 7.62 (d, J = 8.6, 2H), 7.40 (d, J = 6.9, 1H), 7.19–7.13 (m, 1H), 6.96 (td, J = 8.5, 2.5, 1H), 5.73 (s, 1H), 4.58 (s, 2H), 3.84–3.78 (m, 2H), 3.31–3.23 (m, 3H), 2.87 (d, J = 13.8, 1H), 2.69 (d, J = 13.9, 1H), 2.67–2.61 (m, 2H), 2.47–2.32 (m, 4H). 13C NMR (176 MHz, DMSO-d6) δ 168.18, 167.31, 165.80, 162.12 (dd, J = 245.7, 12.4), 159.40 (dd, J = 246.9, 12.2), 150.94, 145.38, 135.71, 132.33, 132.20, 131.96, 130.26, 129.95, 126.52 (d, J = 11.6), 122.59, 111.21 (d, J = 20.3), 104.22, 75.18 (d, J = 4.7), 63.87, 56.02, 54.61, 54.27, 45.27, 41.45, 40.48, 38.45, 30.53. LC/MS (ESI): m/z 617.2. Calculated for C28H27ClF2N6O4S [M + H]+ 617.2.
(Z)-3-(3-(4-(2-(2,4-Difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-3-oxopropyl)-5-(4-methoxybenzylidene)thiazolidine-2,4-dione (50c) was obtained as described above from potassium salt of 5-(4-methoxybenzylidene)thiazolidine-2,4-dione (5c) and 49. Yield 10%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ 8.29 (s, 1H), 7.88 (s, 1H), 7.75 (s, 2H), 7.60 (d, J = 8.8, 2H), 7.57 (d, J = 8.8, 2H), 7.43–7.37 (m, 1H), 7.19–7.14 (m, 1H), 7.11 (t, J = 8.2, 4H), 6.96 (td, J = 8.5, 2.5, 1H), 4.58 (s, 2H), 3.83 (d, J = 4.3, 7H), 3.82–3.79 (m, 1H), 3.30–3.24 (m, 1H), 2.86 (d, J = 14.1, 1H), 2.69 (d, J = 13.9, 1H), 2.66–2.59 (m, 1H), 2.46–2.32 (m, 4H). 13C NMR (176 MHz, DMSO-d6) δ 168.58, 168.21, 167.63, 166.03, 162.12 (dd, J = 245.3, 12.1), 161.65, 161.42, 159.40 (dd, J = 246.9, 12.3), 150.94, 145.37, 133.36, 132.72, 132.53, 132.12, 130.25, 126.53 (d, J = 13.0), 126.02, 125.88, 121.01, 118.48, 115.47, 115.38, 111.21 (d, J = 19.9), 104.22 (t, J = 26.9), 75.18 (d, J = 5.1), 63.88, 55.99, 55.95, 54.62, 54.28, 45.27, 41.45, 40.48, 38.31, 30.62. LC/MS (ESI): m/z 613.2. Calculated for C29H30F2N6O5S [M + H]+ 613.2.
(Z)-5-(2,4-Dichlorobenzylidene)-3-(3-(4-(2-(2,4-difluorophenyl)-2-hydroxy-3-(1H-1,2,4-triazol-1-yl)propyl)piperazin-1-yl)-3-oxopropyl)thiazolidine-2,4-dione (50g) was obtained as described above from potassium salt of 5-(2,4-dichlorobenzylidene)thiazolidine-2,4-dione (5g) and 49. Yield 20%, white crystals. 1H NMR (700 MHz, DMSO-d6) δ = 8.32 (s, 1H), 7.95 (s, 1H), 7.86 (d, J = 2.1, 1H), 7.76 (s, 1H), 7.62 (dd, J = 8.4, 2.0, 1H), 7.58 (d, J = 8.4, 1H), 7.41 (d, J = 7.1, 1H), 7.20–7.14 (m, 1H), 6.98 (s, 1H), 4.59 (s, 2H), 3.85–3.77 (m, 2H), 2.89 (s, 1H), 2.74 (s, 1H), 2.66 (t, J = 7.6, 2H). 13C NMR (176 MHz, DMSO-d6) δ 168.20, 167.09, 165.45, 164.48, 162.77, 162.20 (dd, J = 246.0, 12.3), 159.39 (dd, J = 247.0, 12.2), 150.98, 145.43, 136.17, 135.86, 130.54, 130.47, 130.44, 130.39, 130.26, 128.89, 128.80, 128.50, 127.81, 127.16, 126.19, 125.91, 111.30 (d, J = 19.8), 104.30 (t, J = 26.9), 74.97, 63.64, 55.99, 54.48, 54.18, 40.47, 38.54, 36.25, 31.23, 30.41. LC/MS (ESI): m/z 651.1. Calculated for C28H26Cl2F2N6O4S [M + H]+ 651.1.